Zhang Yongli, Huang Ling, Bi Huichang, Cui Yuqiang, Li Jingqing, Wang Xiangsheng, Qin Xiaoling, Chen Jiangying, Huang Min
Department of Biology, School of Basic Courses, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, University City of Guangzhou, Guangzhou, 510006, Guangdong, People's Republic of China,
Eur J Drug Metab Pharmacokinet. 2013 Jun;38(2):105-13. doi: 10.1007/s13318-012-0102-0. Epub 2012 Jul 14.
Astragalus injection (AI) and Astragalus granules (AG) are the two representative clinical preparations from Astragali Radix. In order to investigate the regulation of metabolism, AI and AG were tested for their ability to affect the major enzyme cytochrome P450 3A isoforms in vivo and in vitro. In the study of CYP3A1 enzyme activity, male rats were pretreated with AI and AG. The "cocktail" approach-based LC-MS/MS results showed that AI pretreatment at 0.16, 0.8 and 4 g kg(-1) day(-1) significantly increased the rat liver microsome CYP3A1 activity by 1.62-, 1.68- and 2.00-fold, and AG pretreatment at 32, 160 and 800 mg kg(-1) day(-1) significantly increased the rat CYP3A1 activity by 1.86-, 2.16- and 1.76-fold. The effects of AI and AG on liver microsome CYP3A1 mRNA expression in rats were analyzed using real-time PCR technique. The results showed that AI and AG pretreatments significantly increased the CYP3A1 mRNA expression. The induction of CYP3A4 enzyme activity by AI and AG in vitro was measured using a CYP3A4 luciferase reporter gene assay in transiently transfected human intestinal LS174T cells. Compared to the control group, AI at 62.5-1,000 mg/ml could significantly induce CYP3A4 reporter gene luciferase activity of 1.36- to 1.88-fold for 48-h incubated PXR-transfected LS174T cells, and AG at 62.5-1,000 μg/ml significantly transactivated CYP3A4 reporter gene luciferase activity of 1.36- to 2.05-fold. However, the CYP3A4 reporter gene construct was not significantly transactivated by the AI and AG in CAR-transfected LS174T cells. These CYP3A isoforms upregulation results can help us to use AI and AG rationally in the clinic.
黄芪注射液（AI）和黄芪颗粒（AG）是黄芪的两种代表性临床制剂。为了研究其对代谢的调节作用，对AI和AG在体内和体外影响主要酶细胞色素P450 3A亚型的能力进行了测试。在CYP3A1酶活性研究中，对雄性大鼠进行AI和AG预处理。基于“鸡尾酒”法的液相色谱-串联质谱结果显示，AI以0.16、0.8和4 g kg⁻¹ day⁻¹预处理可使大鼠肝微粒体CYP3A1活性分别显著提高1.62倍、1.68倍和2.00倍，AG以32、160和800 mg kg⁻¹ day⁻¹预处理可使大鼠CYP3A1活性分别显著提高1.86倍、2.16倍和1.76倍。采用实时PCR技术分析AI和AG对大鼠肝微粒体CYP3A1 mRNA表达的影响。结果显示，AI和AG预处理均显著增加CYP3A1 mRNA表达。在瞬时转染人肠LS174T细胞中，采用CYP3A4荧光素酶报告基因检测法测定AI和AG在体外对CYP3A4酶活性的诱导作用。与对照组相比，AI在62.5 - 1000 mg/ml时，对于经孕烷X受体（PXR）转染的LS174T细胞孵育48小时，可显著诱导CYP3A4报告基因荧光素酶活性提高1.36至1.88倍，AG在62.5 - 1000 μg/ml时可显著激活CYP3A4报告基因荧光素酶活性提高1.36至2.05倍。然而，在经组成型雄烷受体（CAR）转染的LS174T细胞中，AI和AG对CYP3A4报告基因构建体无显著激活作用。这些CYP3A亚型上调结果有助于我们在临床上合理使用AI和AG。
